Molecular expression analysis and characterization of rockfish (Sebastes schlegelii) B cell activating factor

Comp Biochem Physiol B Biochem Mol Biol. 2020 Dec:250:110480. doi: 10.1016/j.cbpb.2020.110480. Epub 2020 Jul 29.

Abstract

B cell activating factor (BAFF) is recognized as a member of the TNF superfamily proteins that mediate the immune responses. In this study, BAFF from rockfish (Sebastes schlegelii) (SsBAFF) was characterized based on its functional aspects. The open reading frame of SsBAFF is 804 bp in length and encodes a 267 long amino acid residue protein with predicted molecular weight of 29.48 kDa. The deduced protein sequence comprises with transmembrane domain, furin cleavage site and TNF domain carrying Flap binding site that unique to TNF family. Recombinant SsBAFF (rSsBAFF) significantly enhanced rockfish lymphocytes proliferation and viability in a concentration dependent-manner according to the results from water soluble tetrazolium salt (WST-1) assay and flow cytometric assay. rSsBAFF also modulated the expression of genes involved in anti-inflammatory (IL-10 and NFκB-2) and anti-apoptotic (Bcl-2 and Bax) signal pathways. SsBAFF mRNA expression was detected ubiquitously in all analyzed rockfish tissues, with the highest levels in the spleen and head kidney. Further, the expression of SsBAFF in spleen were significantly induced following LPS, poly (I:C) and Streptococcus iniae challenges. These findings strongly suggest that SsBAFF might play an important role in rockfish immune system through regulating the inflammatory response and proliferation of immune cells.

Keywords: Cytokine; Lymphocytes; Proliferation; TNF superfamily proteins; Transmembrane domain.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • B-Cell Activating Factor / chemistry
  • B-Cell Activating Factor / genetics*
  • B-Cell Activating Factor / metabolism
  • B-Cell Activating Factor / pharmacology
  • Base Sequence
  • Fish Proteins / chemistry
  • Fish Proteins / genetics*
  • Fish Proteins / metabolism
  • Fish Proteins / pharmacology
  • Gene Expression Regulation*
  • Lymphocytes / cytology
  • Lymphocytes / drug effects
  • Perciformes / genetics*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Recombinant Proteins / pharmacology

Substances

  • B-Cell Activating Factor
  • Fish Proteins
  • Recombinant Proteins